胰岛素样生长因子-1对心肌细胞缺氧损伤的保护作用及机制  

Protective effect and mechanism of insulin-like growth factor-1 on hypoxic injury of cardiomyocyte

作　　者：薛婷匀 李广妹 赵佳叶 孙启天 孙启玉[1] XUE Tingyun;LI Guangmei;ZHAO Jiaye;SUN Qitian;SUN Qiyu(Department of Clinical Laboratory,Affiliated Hospital of Chengde Medical Univercity/Hebei Key Laboratory of Panvascular Diseases,Chengde,Hebei 067000,China;Department of Endocrinology,Affiliated Hospital of Chengde Medical Univercity,Chengde,Hebei 067000,China)

机构地区：[1]承德医学院附属医院检验科/河北省泛血管重点实验室,河北承德067000 [2]承德医学院附属医院内分泌科,河北承德067000

出　　处：《国际检验医学杂志》2024年第19期2323-2328,共6页International Journal of Laboratory Medicine

基　　金：河北省自然科学基金项目(H2022406047)。

摘　　要：目的探讨胰岛素样生长因子-1(IGF-1)预处理对H9c2大鼠心肌细胞缺氧损伤的保护作用,并分析其机制。方法将H9c2细胞随机分成4组,对照组、缺氧组(CoCl2组)、缺氧+IGF-1预处理组(CoCl2+IGF-1组)、缺氧+IGF-1预处理+磷脂酰肌醇3激酶(PI3K)/蛋白激酶B(Akt)通路特异性抑制剂LY294002组(CoCl2+IGF-1+LY294002组)。分别采用CCK8法检测H9c2细胞存活率,Tunel法检测H9c2细胞的凋亡情况,DCFH-DA探针法检测各组H9c2细胞的活性氧(ROS)水平,试剂盒检测细胞超氧化物歧化酶(SOD)、培养基上清乳酸脱氢酶(LDH)活性,酶联免疫吸附试验(ELISA)检测细胞谷胱甘肽过氧化物酶(GSH-Px)及培养基上清丙二醛(MDA)水平,蛋白质印迹法检测H9c2细胞促凋亡蛋白B细胞淋巴瘤2(Bcl-2)关联X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3),抗凋亡蛋白Bcl-2及通路蛋白Akt、磷酸化Akt(p-Akt)表达情况。结果IGF-1能增加细胞活力(P<0.05),降低细胞凋亡率(P<0.05),降低ROS水平(P<0.05),减少MDA产生(P<0.05),降低LDH活性(P<0.05),增加SOD与GSH-Px活性(P<0.05),促进p-Akt表达(P<0.05),减少促凋亡蛋白Bax、Caspase-3表达(P<0.05),增加抗凋亡蛋白Bcl-2表达(P<0.05)。加入PI3K通路特异性阻滞剂LY294002后,IGF-1对缺氧H9c2细胞保护作用消失。结论IGF-1保护心肌细胞免受缺氧造成的损伤,机制是通过激活PI3K/Akt通路抑制心肌细胞氧化应激与凋亡,提高心肌细胞存活率。Objective To investigate the protective effect of insulin-like growth factor-1(IGF-1)preconditioning on hypoxic injury in H9c2 rat cardiomyocytes and its mechanism.Methods H9c2 cells were randomly divided into four groups,control group,hypoxia group(CoCl2 group),hypoxia+IGF-1 pretreatment group(CoCl2+IGF-1 group),hypoxia+IGF-1 pretreatment+phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)pathway specific inhibitor LY294002 group(CoCl2+IGF-1+LY294002 group).CCK8 assay was used to detect the survival rate of H9c2 cells,and Tunel assay was used to detect the apoptosis of H9c2 cells,DCFH-DA probe method was used to detect the reactive oxygen species(ROS)level of H9c2 cells in each group.The activities of superoxide dismutase(SOD)and lactate dehydrogenase(LDH)in culture supernatant were detected by kit.The levels of glutathione peroxidase(GSH-Px)and malondialdehyde(MDA)in culture supernatant were detected by enzyme-linked immunosorbent assay(ELISA).Western blot was used to detect the expression of pro-apoptotic protein B cell lymphoma 2 associated X protein(Bax),Caspase-3,anti-apoptotic protein Bcl-2,pathway proteins Akt and phosphorylated Akt(p-Akt)in H9c2 cells.Results IGF-1 could increase cell viability(P<0.05),reduced cell apoptosis rate(P<0.05),reduced ROS level(P<0.05),re-duced MDA production(P<0.05),reduced LDH activity(P<0.05),and increased SOD and GSH-Px activities(P<0.05).It also promoted the expression of p-Akt(P<0.05),reduced the expression of pro-apoptotic proteins Bax and Caspase-3(P<0.05),and increased the expression of anti-apoptotic protein Bcl-2(P<0.05).After the addition of PI3K pathway specific inhibitor LY294002,the protective effect of IGF-1 on hypoxic H9c2 cells disappeared.Conclusion IGF-1 protects cardiomyocytes from hypoxia injury by inhibiting oxidative stress and apoptosis of cardiomyocytes through activating PI3K/Akt pathway and improving the survival rate of cardiomyocytes.

关 键 词：胰岛素样生长因子-1 缺氧 氧化应激 凋亡 磷脂酰肌醇3激酶/蛋白激酶B通路 

分 类 号：R446.1[医药卫生—诊断学]